CDNA CLONING, SEQUENCING AND EXPRESSION OF HUMAN ETS-1 AND ETS-2

禽成红细胞增多症病毒E26是一种复制缺陷型病毒， 引起鸡成红细胞增多症和成髓细胞增多症的逆转录病毒。 E26的转化基因包括来自两个原癌基因的元件， 鸡的原myb和鸡的原ets，以及病毒性呕吐物中的Deltagag 基因 与E26病毒ets区域同源的人类基因组克隆 被分子克隆并显示与v-ets区域密切相关 通过杂交和部分序列分析。 人类的外星生物DNA 位于两条不同的染色体上 人类染色体上的ets-1位点 11编码一个6.8kb的mRNA，第二个ets-2位点位于21号染色体上 编码4.7、3.2和2.7kb的三种mRNAS。 为了研究 人ets-1和ets-2的结构组织和剪接机制 基因，从人科洛320细胞系制备cDNA文库， 表达非常高水平的ETS特异性转录物。 几 分离与ETS-1和ETS-2探针反应的重组克隆。 这些cDNA克隆正在通过限制性酶切图谱进行表征， Southern印迹分析不同家族的mRNA。 这些克隆 也在测序中。 有初步证据表明， 就ets-1和ets-2而言， 通过选择性剪接事件产生。 全长cDNA克隆 ets-1和ets-2的基因在E.杆菌 和哺乳动物细胞中。 通过上述研究，有可能 精确定义病毒基因及其之间的任何微小差异 细胞对应物--这些差异可能对调节 “基因”的表达或其生物学功能的范围。 这些在进化过程中保守的调控基因， 在细胞分化和增殖中有一定作用。

The avain erythroblastosis virus, E26, is a replication-defective retrovirus which causes erythroblastosis and myeloblastosis in chickens. The transforming gene of E26 includes elements from two proto-onc genes, chicken proto-myb and chicken proto-ets, and Deltagag from the viral gag gene. Human genomic clones homologous to the ets region of the E26 virus were molecularly cloned and shown to be closely related to the v-ets region by hybridization and partial sequence analysis. The human ets DNA is located on two different chromosomes. The human ets-1 locus on chromosome 11 encodes a single mRNA of 6.8 kb; the second ets-2 locus on chromosome 21 encodes three mRNAS of 4.7, 3.2 and 2.7 kb. In order to study the structural organization and splicing mechanism of the human ets-1 and ets-2 genes, a cDNA library was prepared from a human COLO 320 cell line which expresses very high levels of ets-specific transcripts. Several recombinant clones reactive with ets-1 and ets-2 probes were isolated. These cDNA clones are being characterized by restriction mapping and Southern blot analysis into a different family of mRNAs. These clones are also being sequenced. There is preliminary evidence to state that these multiple transcripts, in the case of ets-1 and ets-2, may have been generated through alternative splicing events. The full legnth cDNA clones of ets-1 and ets-2 are being expressed in vitro and in in vivo in E. coli and in mammalian cells. With the above study, it may become possible to define precisely any small differences between the viral genes and their cellular counterparts--differences that may be crucial for the regulation of the "genes" expression or the range of their biological functions. These regulatory genes, which have been conserved during evolution, may have some role in cell differentiation and multiplication.